Yeast translation elongation factor eEF3 promotes late stages of tRNA translocation

Namit Ranjan, Agnieszka A. Pochopien, Colin Chih-Chien Wu, Bertrand Beckert, Sandra Blanchet, Rachel Green, Marina V Rodnina, Daniel N. Wilson

Research output: Contribution to journalArticlepeer-review

Abstract

In addition to the conserved translation elongation factors eEF1A and eEF2, fungi require a third essential elongation factor, eEF3. While eEF3 has been implicated in tRNA binding and release at the ribosomal A and E sites, its exact mechanism of action is unclear. Here, we show that eEF3 acts at the mRNA–tRNA translocation step by promoting the dissociation of the tRNA from the E site, but independent of aminoacyl-tRNA recruitment to the A site. Depletion of eEF3 in vivo leads to a general slowdown in translation elongation due to accumulation of ribosomes with an occupied A site. Cryo-EM analysis of native eEF3-ribosome complexes shows that eEF3 facilitates late steps of translocation by favoring non-rotated ribosomal states, as well as by opening the L1 stalk to release the E-site tRNA. Additionally, our analysis provides structural insights into novel translation elongation states, enabling presentation of a revised yeast translation elongation cycle.

Original languageEnglish (US)
Article numbere106449
JournalEMBO Journal
Volume40
Issue number6
DOIs
StatePublished - Mar 15 2021

Keywords

  • ABC ATPase
  • E-site tRNA
  • L1 stalk
  • cryo-EM
  • eEF3

ASJC Scopus subject areas

  • Neuroscience(all)
  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

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